1121例慢性乙型肝炎患者HBV反转录酶区的耐药突变分析

摘要

Objective To analyze hepatitis B virus (HBV) mutation associated with nucleos(t)ide analogue resistance in multiple sites in the reverse transcriptase (RT) gene in a large sample of patients with chronic hepatitis B. Methods Serum HBV DNA was extracted individually from 1 121 patients. A nested PCR assay with high sensitivity and specifidty developed by our laboratory was employed for the RT gene amplification. Direct aequencing of PCRproduct was performed and the mutations associated with drug resistance at 12 sites were analyzed. The significance of detected mutations was analyzed in collaboration with clinical data of the patients. Results Drug-resistant mutations were detected in 228 patients for lamivudine (LAM), 32 for addfovir (ADV), 13 for entecavir (ETV), and 4 for telbivudine (L-dT). Five mutations were multi-drug resistant. M204V and M204I were dominant LAM-resistant mutations. The former was usually accompanied with L180M mutation, while the latter often emerged alone. N236T±A181 substitution was dominant in ADV-resistant mutation pattern. ETV-resistant mutations occurred on top of LAM-resistant mutations, and T184 exchange was the most common pattern. L-dT resistance was found be M204L Conclusions Detection of HBV drug-resistant mutations at multiple sites of the viral RT gene by direct sequencing is valuable for discovering genotypie drug resistance, thus it is helpful for physicians to adopt anti-HBV therapy more reasonably.%目的 分析大样本慢性乙型肝炎患者HBV反转录酶基因上的多位点核苷(酸)类似物的耐药相关突变情况及其临床意义.方法 提取1 121例患者血清HBV DNA,采用巢式PCR方法 扩增HBV反转录酶(RT)基因,对PCR产物进行DNA双向测序,对12个位点上的耐药相关突变进行检测,并结合临床资料进行分析.结果 检出拉米夫定(LAM)耐药突变228例,阿德福韦(ADV)耐药突变32例,恩替卡韦(ETV)耐药突变13例,替比夫定(L-dT)耐药突变4例.多药耐药5例.LAM耐药突变中以M204V和M2041最常见,前者通常伴随U80M突变,后者常单独出现;ADV耐药突变中以N236T±A181位碱基替换为主;ETV耐药突变发生在LAM耐药基础上,以T184位碱基替换为主;L-dT的耐药突变为M2041.结论 用基因序列测定法检测HBV RT基因多位点耐药相关突变,有助于临床及时发现乙肝患者是否存在HBV基因耐药,合理进行抗病毒治疗.